“If mice were only people….” Re-thinking Pre-Clinical In Vivo Models To Increase the

Probability of Clinical Success

Beth  Ann  Murphy,  PhD  Nina  Jochnowitz  

     

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Pharma  Industry  is  Challenged

2  

ACME    PHARMA  Co.    

Research  Budget  

Scien9fic  and  Technological    Advancements  

•  Many  analyses  that  probe  and  propose  reasons  for  the  decrease  in  R&D  producAvity.  

•  Consensus  is  that  drug  discovery  needs  to  change  to  be  able  to  deliver  novel  drugs  in  the  current  environment.  

Target    Discovery    

Lead  Discovery  

Iden9fy  a  cellular  or  molecular  structure  

involved  in  the  pathology  of  interest.  

Can  you  make  a  drug  to  modulate  the  target?  

Lead  Op9miza9on  Make  the  Lead  have  druglike  proper9es  

Pre-­‐clinical    Safety/Tox   Ph  I   Ph  II   Ph  III   Ph  IV  

Is  the  Lead  safe  to  give  to  people?  

IND/  Clinical    

Candidate   NDA  

DISCOVERY   DEVELOPMENT  CLINICAL  

Preclinical  Candidate  

◦Healthy  ◦PK  ◦Toxicity  

◦Disease  ◦PK/Toxicity  ◦Efficacy?  

◦Longer  ◦Safety/Efficacy  ◦Risk  vs.  Benefit  ◦Label  info.  

Post-­‐marke9ng  

Drug  Discovery  and  Development  Process  

Druglike  Proper9es  •  Bioavailable  •  PharmacokineAc  (ADME)  •  Maintain  effect  on  the  target  

Purpose:  demonstrate  that  the  clinical  candidate  is  safe  and  has  pharmacological  acAvity  that  jusAfies  tesAng  in  

humans.  

Inves9ga9onal  New  Drug  Applica9on  FDA’s  Sponsor’s  Purpose:  seek  approval  to  market.  

Purpose:  Clinical  data  support  labeling  claims  

New  Drug  Applica9on    

Compound  #  

Cost  

5,000-­‐  10,000   1  

Cost  to  bring  a  drug  to  market  was  $2.56  billion  in  2013  dollars.  (Tu?s  Center  for  the  Study  of  Drug  Development).  

Drug Discovery—Convergence of Disciplines

Patent Law

CombinatorialChemistry

SyntheticChemistry

PhysicalChemistry

Biochemistry

Enzymology

Immunology Pharmacology

InformationTechnology

Modelling

Toxicology

Pathology Behavior

Physiology

Project Management

Medicine

Epidemiology

Genetics

What  is  the  problem?  

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Problem:  Drugs  discovered  in  the  Preclinical  stage  frequently  fail  to  translate  into  clinical  success  (~10%  overall  success).  •  1991-­‐2000  =  11%  success  rate  (Nat.  Rev.  Drug  Discov.    2004  Aug;  3(8):711-­‐715.)  •  2005-­‐2010  =    6%  success  rate  (Nat.  Rev.  Drug  Discov.    2014  Jun;  13(6):419-­‐431.)  

Preclinical    

But  why  doesn’t  the  preclinical  work  translate  to  humans?    

Ph  3  Ph  1   Ph  2  

1990’s    

100   11  

2000’s    

6  

Animals  ≠  Humans  

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•  GeneAc  •  Surgical  ManipulaAon  •  Mimic  a  chronic  condiAon  in  an  acute  Ame-­‐frame.  •  Apply  animal  behavior  to  a  human  behavior.  

=  ≠  

•  Effects  in  preclinical  in  vivo  disease  models  are  poor  predictors  of  efficacy  in  the  clinic.  

Pathological    Process  

PHARMACOKINETICS   PHARMACODYNAMICS  

TransducAon  to  Efficacy/Safety  Target  Exposure  

Target  Occupancy  

kon  

koff  

Target  Engagement  

Target  Mechanism  

Biological  Process  

Outcome  Patho-­‐physiology  

Cp  Drug   Ce  

Plasma  

keo  

Target  site  

Biological    Process  

Think  about  it  Differently:    What  is  a  Drug  Actually  Doing

1.  An  integrated  and  quanAtaAve  understanding  of  the  PKPD  relaAonships  and  how  these  translate  to  humans.    Human  PK  –  can  we  get  enough  drug  to  

where  it  needs  to  be?  Human  Target  Engagement  –  can  we  

modulate  the  target  with  the  right  intensity  and  duraAon?  

 

Think  about  it  Differently  

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PHARMACOKINETICS   PHARMACODYNAMICS  

TransducAon  to  Efficacy/Safety  Target  Exposure  

Target  Occupancy  

kon  

koff  

Target  Engagement  

Target  Mechanism  

Biological  Process  

Outcome  Patho-­‐physiology  

Cp  Dose   Ce  

Plasma  

keo  

Target  site  

2.   Confidence  in  target-­‐disease  linkage.  Human  geneAcs  Validated  preclinical  model  

1.   2.  

3.   Iden9fy  and  test  assump9ons  Integrate  data  to  reduce  

assumpAons  IdenAfy  criAcal  assumpAons  and  

assess  impact    

TranslaBonal  Biomarker  Bubble  Diagram

Proof  of  Concept  Proof  of  Mechanism  Human

Preclinical Species

Patho-physiology or Disease

Process

Efficacy / Outcome

Patho-physiology or Disease

Process

Efficacy/Outcome

Target Mechanism

of Action Biomarker

Physiological ResponseBiomarker

In-Vitro Assays

DrugConcentration

(PK)

In-Vitro Assays

DrugConcentration

(PK)

Target Engagement

Biomarker

Target Mechanism

of Action Biomarker

Physiological ResponseBiomarker

Target Engagement

Biomarker

Example  :  Respond  to  Data  from  a  Clinical  Trial  

LDL   HDL  XCETP  Catalyzes  the  exchange  of  TG  and  CE  between  HDL  and  VLDL/LDL  

CETP  

TG  

CE  

Inhibit  CETP   Increase  HDL     Protect  from  CVD    

“BAD”   “GOOD”  

Increase  HDL     Protect  from  CVD    

Lead-­‐Op9miza9on  Drug  Discovery  Program  for  Cardiovascular  Disease    

What  do  we  need  to  ask?/  What  do  we  know?  /What  do  we  have?

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Commercial  CETP  Inhibitor  Screening  Kit  

Humanà  Yes  Bloodà  Yes  

1.  What  is  the  target?à  CETP    

2.  What  do  we  want  the  drug  to  do?à  Inhibit  CETP    

3.  What  is  the  effect  of  inhibiAng  CETP?à  Increases  HDL    

Able  to  measure  drug  levels  in  the  

circula9on  (Pharmacokine9cs)  

Commercial  HDL  Screening  Kit  

Humanà  Yes  Bloodà  Yes  

CETP  

Human  CETP    Transgenic    Mouse  

   

Long  9me  period  

●Mice  do  not  develop  CVD  naturally.  ●Mice  ≠Humans  

Measure  of  clinical  effect  =  Decreased  CVD  related  deaths    

TranslaBonal  Biomarker  Bubble  Diagram:    CETP  Inhibitor  Program

Proof  of  Concept  Proof  of  Mechanism  Human

CETP Mouse

Patho-physiology or Disease

Process

Efficacy / Outcome

Patho-physiology or Disease

Process

Efficacy/Outcome

Target Mechanism

of Action Biomarker

Physiological ResponseBiomarker

In-Vitro Assays

DrugConcentration

(PK)

In-Vitro Assays

DrugConcentration

(PK)

Target Engagement

Biomarker

Arrow  =  Can  be  measured  both  in  preclinical  experiments  and  in  human  clinical  trials  

Target Mechanism

of Action Biomarker

Physiological ResponseBiomarker

Target Engagement

Biomarker

CETP  inhib.  w/  

Kit  

PK  Assay  

CETP  inhib.  w/  

Kit  

PK  Assay  

CETP  inhib.  From  human  

blood  

CETP  inhib.  From  human  

blood  

HDL  

HDL  

CV  Deaths  

Diabetes,  Athero,  HT,  etc.    

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Case  Study  

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Case  Study  An9-­‐Thrombo9c  Drug  For  Stroke  and  AFIB  

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What are the Main Cause and Effects of Thrombotic Events1.  Cause Atrial Fibrillation 2.  Effect Stroke(s)

What is the Human cost?1.  Lives2.  Annually, Death rate is nearing 300,000 in the

USA3.  More than 1,500,000 hospitalizations annually

What is the Financial cost?1.  Personal2.  Broad Economic losses (work related)3.  Forty Billion ($40,000,000) dollars!

Atrial Fibrillation - AFIBDisease of the heart characterized by irregular and often faster heartbeat.Causes of an Irregular Heartbeat•  Hypertension, Diabetes, Congestive heart failure, Dehydration,

Hyperkalemia, Mitro-valve Prolapse, Poisoning, (cocaine, amphetamine, digitalis…), Anaphylaxis, hyper & hypo-thyroidism, Cardiomyopathy

Indications•  Abnormal electrical discharges (signals) that generate chaotically

throughout the upper chambers of the heart (atria).•  Reduction in the Atria to pump blood into the ventricles

•  Response is the heart to beat too rapidly.•  AFIB causes turbulence of the blood which causes clot formation.•  https://youtu.be/fxUITWjrhhs• Management: Rate Control, Maintenance of normal rhythm, stroke

preventionCost

•  Prevention: Pharmacological and/or surgical (Cardio conversion, Catheter Ablation, surgical ablation, atrial pacemaker)

•  Annually more than 130,000 deaths in the USA. •  More than 750,000 hospitalizations occur annually. •  AFib costs the United States about $6 billion each year, diagnosis and

treatment. 16  

StrokesSudden death of brain cells from lack of oxygen.

•  Caused by blockage of blood flow or rupture of an artery to the brain.•  Ischemic stroke (part of the brain loses blood flow, bleeding from periphery)•  Hemorrhagic stroke (brain bleed, Transient Ischemic Attack, TIA >24 hours)

Indications•  Sudden loss of speech, weakness, or paralysis of one side of the body can

be symptoms. •  Confirmation by scanning the brain with special X-ray tests, such as CAT

scans.Costs

The death rate and level of disability resulting from strokes can be dramatically reduced by immediate and appropriate medical care. •  Prevention involves minimizing risk factors, such as controlling high blood

pressure and diabetes. •  Stroke kills about 140,000 Americans each year—that’s 1 out of every 20

deaths.•  Annually795,000 people in the United States have a stroke. About 610,000

of these are first or new strokes•  About 87% of all strokes are ischemic strokes.•  Stroke costs the United States an estimated $34 billion 

Animal Models: AV Shunt

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Project  A:  An9-­‐Coagulant  Drug  Discovery  Program:  Intrinsic  Pathway  Inhibitor  

CLOT  18  

ELIQUIS  (apixaban)  PRT-­‐054,021  (betrixaban)  SAVAYSA(edoxaban)  XARELTO(rivoroxaban)  

Vitamin  K    antagonists  (warfarin)  

Vitamin  K    antagonists  (warfarin)  

Coagula9on  Factor  Targets  of    Currently    Approved  SOC  or  An9-­‐Coagulant  Drugs  

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What  do  we  need  to  ask?/  What  do  we  know?  /What  do  we  have?

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1.  What  is  the  target?à  fIXa    

2.  What  do  we  want  the  drug  to  do?à  Inhibit  excess  thromboAc  acAvity    

3.  What  is  the  effect  of  inhibiAng  factor  9?à  Decreased  in  thromboAc  events      

Able  to  measure  drug  levels  in  the  circula9on  (Pharmacokine9cs)  

Commercial  aPTT  Screening  Kit  

Humanà  Yes  Bloodà  Yes  

   

Long  9me  period  

●Rabbits  do  not  develop  CVD  naturally.  ●Rabbits  ≠  Humans  

Measure  of  clinical  effect  =  Decreased  CVD  related  deaths    

AVS  Model  

Commercial  FIXa  Inhibitor  Screening  Kit  

Humanà  Yes  Bloodà  Yes  

AVS  

aPTT  

aPTT:  A  Rou9ne  Clinical  Test  and  a  Biomarker  of    Intrinsic  Pathway  Coagula9on  

 ACTIVATED  PARTIAL    THROMBOPLASTIN  TIME  

(APTT)    21  

TIME  TO  CLOT  

Inhibitors  of  the  Intrinsic  Pathway  à      

↑  APTT  

Back  

Arteriovenous  (AV)  Shunt  in  Rabbits  :      Model  to  assess  an9-­‐coagula9on.  

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To carotid artery To jugular vein

AV-Shunt

ThreadThrombus

AV-shunt

Thread

Thrombus

Femoral A. Femoral V.

•  Method:    –  Connect  an  artery  and  vein  to  a  shunt.    –  A  thrombogenic  sAmulus  (e.g.  thread)  is  inside  the  

shunt  .      –  As  blood  flows  through  the  shunt  à  clot  will  form  on  

the  thread.    –  Weigh  the  clot.  

•  An9-­‐Coagula9on  =  clot  weight,  blood  sampling  

•  Issues:  AVS  is  a  mixed  arterial  venous  anAthromboAc  model.  

•  Needs:  To  enhance  our  confidence  in  the  predicAve  value  of  our  preclinical  models,  translaAonal,  or  creaAng  a  plan  to  further  characterize  acAvity  of  SoC,  define  target  engagement  levels  etc.    

Back  

Arteriovenous  (AV)  Shunt  in  Rats  :      Model  to  assess  an9-­‐coagula9on.  

23  

To carotid artery To jugular vein

AV-Shunt

ThreadThrombus

AV-shunt

Thread

Thrombus

Femoral A. Femoral V.

•  Method:    –  Connect  an  artery  and  vein  to  a  shunt.  –  A  thrombogenic  sAmulus  (e.g.  thread)  is  inside  the  

shunt  .      –  As  blood  flows  through  the  shunt  à  clot  will  form  on  

the  thread.    –  Weigh  the  clot.  

•  An9-­‐Coagula9on  =  clot  weight  •  Can  take  blood  samples  from  the  AVS  rats.  

Work  Sheet

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TranslaBonal  Biomarker  Bubble  Diagram:    

Proof  of  Concept  Proof  of  Mechanism  Human

Model

Patho-physiology or Disease

Process

Efficacy / Outcome

Patho-physiology or Disease

Process

Efficacy/Outcome

Target Mechanism

of Action Biomarker

Physiological ResponseBiomarker

In-Vitro Assays

DrugConcentration

(PK)

In-Vitro Assays

DrugConcentration

(PK)

Target Engagement

Biomarker

Arrow  =  Can  be  measured  both  in  preclinical  experiments  and  in  human  clinical  trials  

Target Mechanism

of Action Biomarker

Physiological ResponseBiomarker

Target Engagement

Biomarker

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In  vitro  assay:  Measure  of  in  vitro  acAvity,  such  as  potency  or  affinity  Drug  concentra9on:  The  pharmacokineAcs  of  the  compound  typically  measured  as  unbound  plasma  concentraAons  and/or  target  site  exposure  Target  engagement  biomarker:  Measurement  of  compound  binding  to  target,  such  as  with  PET  or  measuring  anAbody  bound  to  anAgen.  Target  Mechanism  of  Ac9on  Biomarker:  A  proximal  biochemical  or  proximal  physiological  (eg  electrophysiological)  response  as  a  result  of  compound  interacAon  with  the  target    Physiological  Response  Biomarker:  A  physiological  or  Assue  response  driven  by  compound  acAvity  at  the  target,  but  not  directly  linked  to  pathophysiology  Pathophysiology  or  Disease  Process:  A  biochemical  response  involved  in  the  disease  process  or  acAvity  in  an  animal  model  or  on  a  clinical  endpoint  that  serves  as  an  index  of  the  disease  process  Efficacy/Outcome:  AcAvity  in  an  animal  model  of  disease  that  has  been  demonstrated  to  predict  clinical  efficacy  or  posiAve  effect  on  a  clinical  endpoint  endorsed  by  regulators  as  sufficient  for  approval    

Explana9on  of  Bubbles    

Case  Study:  TranslaBonal  Biomarker  Bubble  Diagram

Proof  of  Concept  Proof  of  Mechanism  

Human

Patho-physiology or Disease

ProcessEfficacy / Outcome

Patho-physiology or Disease

Process

Efficacy/Outcome

In-Vitro Assays

DrugConcentration

(PK)

In-Vitro Assays

DrugConcentration

(PK)

aPTT  

AVS    aPTT,   Ex  vivo  aPTT  

Plasma  

Plasma  

Green  shading  =  Biomarkers  that  are  translaAonal  and/or  support  IVIVC  and  have  been  validated/reduced  to  pracAce  Blue  arrow  =  Need  for  mathemaAcal  translaAonal  modelling  across  species  *  Indicates  assay  in  development  

Target Mechanism

of Action Biomarker

Physiological ResponseBiomarker

Target Mechanism

of Action Biomarker

Physiological ResponseBiomarker

Target Engagement

Biomarker

Ex  vivo  aPTT,  PT  

Phase  II:  ThromboAc  

events;  Stroke,  TIA,  Pulmonary  

Phase  III:CVD  Related  Deaths  

X X

X XRabbit

Target  Engagement  

Target Engagement

BiomarkerX